含RGD修饰的丹参酮ⅡA多级靶向纳米粒的质量评价

目的探讨丹参酮ⅡA多级靶向纳米粒的制备及其质量评价。方法采用乳化溶剂蒸发法制备丹参酮ⅡA多级纳米粒;测定丹参酮ⅡA多级靶向纳米粒的粒径分布及纳米微粒表面结构;并检测丹参酮ⅡA多级靶向纳米粒的载药量、包封率及体外药物释放规律。结果课题组制备的丹参酮ⅡA多级靶向纳米粒,大小均匀,载药纳米粒的平均粒径为190nm,Zeta电位为4.3mV,包封率(94.12±5.20)%,载药量(2.05±0.12)%。与游离的丹参酮ⅡA单体相比,丹参酮ⅡA多级靶向纳米粒释放速度明显减慢,在120h累积释放量为72.59%。结论采用乳化溶剂蒸发法成功制备了含RGD修饰的丹参酮ⅡA多级靶向纳米粒。与丹参酮ⅡA单体相比,制备成纳米制剂后,多级靶向载药纳米粒能随着时间的延长将药物逐步释放出来,具有良好的缓释特征。     

载奥沙利铂的还原敏感型四氧化三铁纳米粒的体内外靶向性评价

化疗药物的非特异性蓄积和释放是影响其治疗效果以及引起不良反应的主要原因。现阶段,将药物纳米制剂化并且进行响应性释药设计是提高药物肿瘤特异性蓄积量和降低其不良反应的重要策略。本研究首先合成了一种α-烯醇化酶靶向肽修饰的共价荷载奥沙利铂前药的聚乙二醇聚赖氨酸嵌段共聚物,通过相转透析法制备了载药聚合物包覆的四氧化三铁纳米粒,以提高奥沙利铂的循环稳定性及肿瘤靶向性。在体外和活体水平对靶向修饰的载药四氧化三铁纳米粒的物理化学性质、还原响应药物释放、细胞摄取和肿瘤靶向等生物学功能进行了相关研究。体外的还原响应释药、肿瘤靶向摄取及摄取抑制考察结果显示,在模拟肿瘤细胞浆微环境的还原条件中,载药纳米粒可实现3 h内超80%的奥沙利铂原型药物的快速释放;流式细胞术的结果显示,靶向多肽的修饰能够增加肿瘤细胞对载药纳米粒的摄取量,并且靶向载药纳米粒主要是通过受体蛋白和小窝蛋白介导的能量依赖的内吞途径被肿瘤细胞所摄取的。所有动物实验操作均通过复旦大学药学院实验动物伦理委员会批准并遵循相关管理规定。药物动力学实验结果显示,纳米制剂化能显著增加奥沙利铂的平均药时曲线下面积(AUC_0-∞),约为...     

抗体偶联药物的有关进展

化疗肿瘤治疗的常规手段之一,但由于细胞毒药物没有明显的靶向性,导致其副作用十分显著。因此免疫偶联药物由于其良好的靶向性及抗癌活性就成为了研究的热点。文章综述了细胞毒药物与单克隆抗体偶联的各种常用方法,包括使用偶联剂进行偶联、利用肽链进行偶联、利用二硫键进行偶联和利用大分子技术进行偶联,以及抗体偶联药物的研究现状。抗体偶联药物有细胞毒药物、抗体和偶联剂组成,利用各种偶联手段将单克隆抗体与细胞毒药物偶联,不仅可以将细胞毒药物特异性地靶向于肿瘤细胞,而且可以降低细胞毒药物对于正常细胞的杀伤,同时还可以具备缓释、提高药物抗肿瘤活性等效果,是未来肿瘤靶向治疗的发展方向之一。同时文章还结合作者的自身工作,对抗体偶联药物的未来发展方向提出了建议。     

聚乳酸及其共聚物脑靶向载药纳米粒的研究进展

目的：综述聚乳酸及其共聚物（PLA/PLGA）脑靶向载药纳米粒（NP）的研究进展。方法：查阅2005-2012年来国内外有关文献,对PLA/PLGA脑靶向载药NP的作用机制、制备方法、功能分子的连接方式、体外体内作用效果评价方法进行综述。结果与结论：受体介导入脑是目前最为成熟的脑靶向给药机制,鼻腔给药也是一种有效的入脑机制;脂溶性载药NP和水溶性载药NP的制备分别采用乳化溶剂挥发法和复乳法;而溶剂扩散法可加快微球的形成,用于制备粒径较小的NP;功能分子主要利用其上的巯基、氨基、羧基或马来酰亚胺等活性基团与PLA或PLGA对应基团进行共价结合,或利用生物素-亲和素体系脑靶向递药;体外效果评价常用大鼠脑微血管内皮细胞（BMVEC）单层培养模型及BMVEC-星形胶质细胞共培养模型;体内效果评价常用荧光显微镜和活体荧光成像、放射自显影法,前者用于考察以荧光素和香豆素等荧光染料标记的脑靶向NP,后者用于以125I、3H、14C等放射性核素标记的脑靶向NP。提高脑靶向的靶向效率、提高载药量、减少PLA/PLGA的用量、改善PLA/PLGA纳米粒体内降解性能和增加稳定性,是今后脑靶向PLA/PLGA载药NP研究的重点方向。     

抗肿瘤药物靶向载体系统的研究与开发

综述肿瘤靶向给药的基础和抗肿瘤药物靶向载体系统的发展。分类介绍普通被动靶向载药系统（如微乳、传统脂质体、聚合物纳米粒、固体脂质纳米粒、纳米脂质载体、药-脂结合物纳米粒等）、表面修饰的被动靶向载药系统及主动靶向载药系统（如免疫脂质体、免疫聚合物纳米粒及受体-配体介导靶向纳米载体）的研究与开发。在传统药物制剂的基础上，发展抗肿瘤药物的新型靶向载体系统，改善药物在体内的代谢动力学特性，增加药物定向富集到肿瘤部位甚至肿瘤细胞内，提高疗效，降低毒副作用，是近年来备受关注的课题。     

贝伐单抗介导的阿霉素白蛋白纳米粒的制备与优化

目的采用Box-Behnken效应面法优化贝伐单抗介导的阿霉素白蛋白纳米粒制备工艺。方法在采用去溶剂化-固化交联法制备出的阿霉素白蛋白纳米粒的基础上,用异型双功能交联剂NHS-PEG3500-MAL作为偶联剂,将贝伐单抗偶联到载药白蛋白纳米粒表面。以2-亚氨基硫烷盐酸盐用量、载药白蛋白纳米粒与贝伐单抗质量比、NHS-PEG3500-MAL用量为影响因素,以纳米粒粒径、载药量和包封率作为评价指标,用三因素三水平的Box-Be-hnken效应面法设计试验,并对免疫纳米粒的制备进行优化;考察制备出的免疫纳米粒的抗体活性保存率及稳定性。结果优化后的处方是2-亚氨基硫烷盐酸盐50μg、DOX-A-NPs与Bevacizumab质量比为27.5 mg.mg-1、NHS-PEG3500-MAL用量为8.8 mg,以此处方制得的免疫纳米粒粒径为(216.1±2.31)nm、载药量为(28.93±0.94)%、包封率为(80.39±2.83)%,测得值与预测值相差较小。结论采用Box-Behnken效应面法优化并制备阿霉素白蛋白免疫纳米粒是可行的。     

阿苯达唑聚氰基丙烯酸正丁酯纳米粒的制备、性质及其组织靶向性研究阿苯达唑聚氰基丙烯酸正丁酯纳米粒的制备、性质及其组织靶向性研究

目的制备阿苯达唑聚氰基丙烯酸正丁酯纳米粒（albendazole polybutycyanocrylate nanoparticles，ABZ-PBCA-NP）TDDS给药系统，并考察相关特性及组织分布靶向性。方法种子乳化聚合法制备阿苯达唑纳米粒；等温吸附法考察纳米粒载药特性；动态透析法研究4种制剂的体外释药动力学；同位素标记阿苯达唑纳米粒在小鼠脏器组织分布和生物利用度。结果ABZ-PBCA-NP体外释药遵循Higuchi方程，加入PVP制成的载药纳米粒符合双指数函数。纳米粒的载药方式遵循Langmuir吸附方程。小鼠ig ³H-ABZ-PBCA-NP后, 药物的肝、脾中的靶向指数分别为11.4和3.9，阿苯达唑纳米粒和混悬剂相对生物利用度分别为76.0%和36.9%。结论制备纳米粒加入PVP可使药物具吸附性和分散性，纳米粒载体可降低药物与血浆蛋白结合率，增强药物的肝、脾脏器靶向性和延缓释药。     

肝靶向纳米粒的研究进展

纳米粒具有靶向性,能直接向靶器官、靶细胞或细胞内靶结构输送药物,同时具有缓释、保护药物、提高疗效、降低毒副作用等优点.肝靶向纳米粒可用于肝肿瘤的化疗、病毒性肝炎的治疗、细胞内感染的治疗、抗肝部寄生虫及基因治疗.随着纳米粒研究的不断深入,在肝肿瘤及非肿瘤领域,纳米粒都将有着极为广阔的发展前景.     

抗癫疒间肽纳米粒的制备及体外释药性能的研究

目的制备抗癫疒间肽纳米粒,并研究其体外释药性能。方法选用聚乙二醇-聚乳酸-聚乙醇酸嵌段共聚物为载体,采用复乳-溶剂挥发法制备抗癫疒间肽纳米粒,以包封率、载药量等指标优化制备工艺,并研究纳米粒体外释药性能。结果抗癫疒间肽纳米粒外观呈圆形或类圆形,平均粒径为(100.2±2.45)nm,包封率和载药量分别为(64.46±1.34)%和(4.73±0.32)%,体外释药呈现缓释和突释两个阶段,符合Weibull方程。结论建立的制备工艺简便可行,得到的抗癫疒间肽纳米粒包封率和载药量较高,粒径小,体外释药具有明显的缓释特征。     

叶酸偶联壳聚糖载多西他赛纳米粒的制备

目的:探索靶向叶酸受体的多西他赛(DTX)纳米粒的制备方法。方法:利用叶酸活性酯与壳聚糖分子上的氨基反应,制得叶酸偶联壳聚糖(FA-CTS);再通过离子交联法,将DTX作为模型药物,制备叶酸偶联壳聚糖载DTX(FA-CTS/DTX)纳米粒。以载药量、包封率、粒径和跨距为指标,采用星点设计-效应面法优化搅拌速率、DTX加入量、壳聚糖-三聚磷酸钠(CTS-STPP)的质量比,并进行验证。利用激光粒度分析仪测定纳米粒粒径大小及分布,在磷酸盐缓冲液中对载药纳米粒进行体外释药试验。结果:最优处方(处方量为2.5 mg)为搅拌速率为1 300 r/min、DTX加入量为0.58μg,CTS-STPP的质量比为5.55。所制备的FA-CTS/DTX纳米粒平均粒径为(232.8±0.43)nm、包封率为(86.74±0.60)%、载药量为(25.29±3.21)%、跨距为0.039±1.02;30 min内累积释药40.22%,随后缓慢释放,24 h内累积释药80.25%。结论:成功制备具有缓释作用的FA-CTS/DTX纳米粒。     

Formulation development of albumin based theragnostic nanoparticles as a potential delivery system for tumor targeting

Background: Generally, chemotherapeutic drugs attack on both normal and tumor cells non-specifically causing life threatening side effects, necessitating targeted drug delivery to tumors.

Purpose: The purpose of this study is to formulate albumin-based nanoparticles for tumor targeted drug delivery and noninvasive diagnosis.

Methods: Albumin based nanoparticles (NPs) were developed as a potential tumor theragnostic agent by entrapping an anti cancer drug, doxorubicin and a near infrared dye, indocyanine green. Theragnostic nanoparticles were prepared using a well established coacervation/nanoprecipitation method followed by lyophilization. The formulation was optimized by varying process parameters using full factorial design of experiments. Release of dye and drug from NPs and physical state of the drug in NPs was studied using DSC. The NPs were injected into tumor bearing mice intravenously and imaged using a bio-imager.

Results: The optimized nanoparticle formulation had a particle size of 125.0?±?1.8?nm, poly dispersity index of 0.180?±?0.057 and zeta potential of ?32.7?±?0.9 mV. The release of dye and drug from the nanoparticles was determined to be quasi-fickian diffusion mediated. Differential scanning calorimetry (DSC) studies revealed the stability of drug in the NP. The in-vivo studies showed enhanced accumulation of the dye loaded NPs at the tumor site than the dye solution, thus allowing noninvasive tumor monitoring.

Conclusion: These results project the newly proposed and evaluated nanoparticle formulation as a potential tumor targeting and imaging delivery system.     

Legumain/pH dual-responsive lytic peptide–paclitaxel conjugate for synergistic cancer therapy

After molecule targeted drug, monoclonal antibody and antibody–drug conjugates (ADCs), peptide–drug conjugates (PDCs) have become the next generation targeted anti-tumor drugs due to its properties of low molecule weight, efficient cell penetration, low immunogenicity, good pharmacokinetic and large-scale synthesis by solid phase synthesis. Herein, we present a lytic peptide PTP7-drug paclitaxel conjugate assembling nanoparticles (named PPP) that can sequentially respond to dual stimuli in the tumor microenvironment, which was designed for passive tumor-targeted delivery and on-demand release of a tumor lytic peptide (PTP-7) as well as a chemotherapeutic agent of paclitaxel (PTX). To achieve this, tumor lytic peptide PTP-7 was connected with polyethylene glycol by a peptide substrate of legumain to serve as hydrophobic segments of nanoparticles to protect the peptide from enzymatic degradation. After that, PTX was connected to the amino group of the polypeptide side chain through an acid-responsive chemical bond (2-propionic-3-methylmaleic anhydride, CDM). Therefore, the nanoparticle (PPP) collapsed when it encountered the weakly acidic tumor microenvironment where PTX molecules fell off, and further triggered the cleavage of the peptide substrate by legumain that is highly expressed in tumor stroma and tumor cell surface. Moreover, PPP presents improved stability, improved drug solubility, prolonged blood circulation and significant inhibition ability on tumor growth, which gives a reasonable strategy to accurately deliver small molecule drugs and active peptides simultaneously to tumor sites.     

载阿霉素透明质酸聚合物纳米粒的构建及其体外性质研究

目的 合成透明质酸（HA）接枝单油酸甘油酯（GMO）两亲性聚合物HGO,并研究其所制备载阿霉素（DOX）纳米粒的理化性质及体外抗肿瘤效果。方法 HA与GMO通过酯化反应制得载体聚合物HGO,通过核磁共振波谱法及红外光谱法对其进行结构表征;采用芘荧光探针法测定聚合物临界聚集浓度（CAC）。采用透析法制备聚合物HGO载阿霉素（DOX@HGO）纳米粒,并对其进行粒径分布、Zeta电位及微观形态的表征;通过检测其在不同离子强度、不同pH条件下的粒径变化考察纳米粒的体外稳定性;考察DOX@HGO纳米粒在不同pH条件下的体外释放行为;CCK-8法考察DOX@HGO纳米粒对MDA-MB-231细胞的体外抑瘤效果;并通过荧光显微镜研究MDA-MB-231细胞对DOX溶液、DOX@HGO纳米粒的摄取能力,以及HA预处理对DOX@HGO纳米粒摄取的影响。结果 成功制得两亲性聚合物HGO,聚合物HGO中GMO的取代度为15.8%,CAC为0.023 mg·mL^-1。DOX@HGO纳米粒呈规则的球形,平均粒径为（130.800±1.709）nm,平均电位为（-32.600±0.153）mV,包封率和载药量分别为（98.65±0.74）%和（33.03±0.17）%,在不同离子强度下、模拟胃肠液中表现出良好的稳定性;DOX@HGO纳米粒的体外释放表现出pH依赖性。体外抗肿瘤活性实验表明,DOX@HGO纳米粒对MDA-MB-231细胞的生长具有较好的抑制作用;与DOX溶液比较,DOX@HGO纳米粒显著增加肿瘤细胞对于DOX的摄取（P＜0.05） ,HA预处理显著减少肿瘤细胞对DOX@HGO的摄取（P＜0.05）。结论 所构建的DOX@HGO纳米粒具有良好的理化性质,并且具有一定的pH敏感性及靶向抗肿瘤细胞的能力,是具有应用潜力的药物载体。     

Anti-P-glycoprotein conjugated nanoparticles for targeting drug delivery in cancer treatment

Targeting therapeutics to specific sites can enhance the efficacy of drugs, reduce required doses as well as unwanted side effects. In this work, using the advantages of the specific affinity of an immobilized antibody to membrane P-gp in two different nanoparticle formulations were thus developed for targeted drug delivery to multi-drug resistant cervical carcinoma (KB-V1) cells. Further, this was compared to the human drug sensitive cervical carcinoma cell line (KB-3-1) cells. The two nanoparticle preparations were: NP1, anti-P-gp conjugated with poly (DL-lactic-coglycolic acid) (PLGA) nanoparticle and polyethylene glycol (PEG); NP2, anti-P-gp conjugated to a modified poloxamer on PLGA nanoparticles. The cellular uptake capacity of nanoparticles was confirmed by fluorescent microscopy. Comparing with each counterpart core particles, there was a higher fluorescence intensity of the targeted nanoparticles in KBV1 cells compared to KB-3-1 cells suggesting that the targeted nanoparticles were internalized into KB-V1 cells to a greater extent than KB-3-1 cell. The results had confirmed the specificity and the potential of the developed targeted delivery system for overcoming multi-drug resistance induced by overexpression of P-gp on the cell membrane.     

肝癌特异性多柔比星免疫磷脂纳米粒的实验研究

目的:研制肝癌特异性多柔比星免疫磷脂纳米粒。方法:先制备出多柔比星聚氰基丙烯酸正丁酯毫微粒,再采用磷脂双分子层包裹,得到多柔比星磷脂纳米粒;将抗酸性铁蛋白单克隆抗体(McAb-PAF)与其相联,最后得到肝癌特异性多柔比星免疫磷脂纳米粒。并对其制备工艺、特性、细胞毒性、体内靶向性等进行了探讨。结果:制得的多柔比星免疫磷脂纳米粒粒径为97.2nm,抗体结合率78.8％,抗体活性保持良好;多柔比星免疫磷脂纳米粒对肝癌细胞的杀伤作用明显强于游离的多柔比星和普通脂质体;对于肝癌细胞荷瘤裸鼠,免疫磷脂纳米粒组较普通脂质体组抑瘤率有显著性差异。结论:肝癌特异性多柔比星免疫磷脂纳米粒,有可能成为一种新的药物靶向载体系统。     

PCL-PEG-PCL载姜黄素纳米粒子的制备以及体外药物释放的考察

目的制备一种生物可降解、生物相容性良好的姜黄素纳米粒子,并对其体外药物释放行为进行考察。方法采用开环聚合法制备生物可降解的PCL-PEG-PCL三嵌段聚合物,然后采用乳液挥发法制备负载姜黄素的PCL-PEG-PCL纳米粒子,通过透射电镜观察所制备纳米粒子的形貌特征,动态光散射(DLS)测定粒径,采用HPLC测定纳米粒子的包封率和载药量,同时考察其体外药物释放行为。结果姜黄素纳米粒子具有球形结构,粒径在200 nm左右,载药量为(14.23±0.35)%,3 d体外累积释药量65%。结论所制备的姜黄素纳米粒子具有较高的载药量和包封率,同时体外药物释放实验证实姜黄素纳米粒子具有良好的缓释功能。     

Combination therapy of cRGD-DOX self-assembled nanoparticles and bevacizumab for breast cancer

The interplay among diverse cell populations in the tumor microenvironment contributes to tumor progression. Targeting to different cell populations might result in improved therapeutic effects on malignant tumors. Integrins high express on many kinds of tumor cells, and VEGF has a strong effect on tumor angiogenesis. Therefore, based on tumor cells and angiogenesis, we fabricated integrin-targeting cRGD-DOX nanoparticles and combined them with the anti-VEGF antibody bevacizumab. We evaluated the antitumor effect of this combination therapy in an integrin-overexpressing MDA-MB-231 tumor model. The cRGD-DOX nanoparticles were effectively uptake by MDA-MB-231 cells and the uptake was related to the expression of integrinin; cRGD-DOX nanoparticles showed less cytotoxic than free DOX; Bevacizumab did not show significant cytotoxicity against MDA-MB-231 cells at concentrations less than 1 mg/mL. The in vivo results showed that bevacizumab could reduce tumor interstitial fluid pressure; the combination of bevacizumab and cRGD-DOX nanoparticles showed enhanced antitumor effects compared with the corresponding single-agent treatments. These findings suggested the combination of angiogenesis antibody and integrin-targeting nanoparticle loaded with a cytotoxic drug was a promising cancer treatment regimen.     

包载尼莫地平的聚乳酸-聚乙二醇嵌段共聚物纳米粒的制备和质量评价

目的制备尼莫地平纳米粒的粉针剂，以期改善尼莫地平难溶于水而导致易析出晶体的问题以及其制剂中乙醇引起血管刺激的毒副作用。方法以聚乳酸、聚乙二醇嵌段共聚物作为载体，用溶剂挥发法制备载药纳米粒水分散体系及其冻干粉针．并评价其粒径、Zeta电位、包封率、载药量、形态和冻干粉针的再分散性等性能。结果制备了136nm载药纳米粒的冻干粉针，Zeta电位为-29．90mV，包封率为67．9％，载药量为6．17％，冻干再分散情况良好。结论所制备的尼莫地平纳米粒基本解决了原有制剂药物溶解性差，易析出晶体的问题，避免使用有机溶剂和有副作用表面活性剂的应用。     

Chitosan nanoparticles for controlled delivery of brimonidine tartrate to the ocular membrane

Various efforts have been made to improve the bioavailability and to prolong the residence time of eye drops. Drug loaded polymeric nanoparticles offer several favorable biological properties. Thus, brimonidine tartrate (BT) loaded chitosan (CS) nanoparticles were prepared by inducing the ionic gelation upon addition of sodium tripolyphosphate (TPP). Nanoparticles were characterized by TEM, SEM, particle size, polydispersity index (PI), DSC, IR, entrapment efficiency which gave an insight of physicochemical interaction that influenced the CS nanoparticle formation and entrapment of BT. In vitro release of BT nanoparticle showed sustained release over the period of 4 h in saline phosphate buffer pH 7.4. Both placebo and BT loaded nanoparticles had a mean particle size range of about 270-370 nm with PI less than 0.5. DSC studies demonstrated structural interactions between BT, TPP and CS matrix. Entrapment efficiency of the CS nanoparticles ranged from 36-49% depending on the CS:TPP weight ratio. In vivo studies confirmed a significant sustained effect of BT nanoparticles compared to conventional eye drops. These results suggest that BT loaded CS nanoparticles could help to reduce dosage frequency by sustained drug release in the treatment of glaucoma.     

Pharmaceutics: Characterization and Trypanocidal Activity of Nifurtimox-containing and Empty Nanoparticles of Polyethylcyanoacrylates

The aim of this study was to evaluate the utility of nanoparticles of polyalkylcyanoacrylate as a targeted delivery system for nifurtimox against Trypanosoma cruzi, responsible for Chagas' disease. Ethylcyanoacrylate nanoparticles were prepared by an emulsion polymerization process and formulations containing different concentrations of nifurtimox, polyethylcyanoacrylates and surfactants were investigated and analysed for size and drug content. The nanoparticles obtained were less than 200 nm in size, as measured by electron microscopy and cytometry. The peak percentage of nifurtimox uptake into the nanoparticles was 33.4% for use of 500 μL polyethylcyanoacrylate, 200 μL surfactant (Tween 20) and 10 mg nifurtimox in 50 mL polymerization medium. The highest release of nifurtimox from the nanoparticles was 65.4% after 6-h incubation at pH 7.4. In-vitro studies using cultures of T. cruzi epimastigotes revealed considerably increased trypanocidal activity compared with a standard solution of nifurtimox. Studies of cell cultures previously infected with metacyclic forms of the parasite showed that only 2-h treatment with solutions of 0.001% of the nanoparticle suspension reduced parasitism by 87–94% both when the nanoparticles were loaded with nifurtimox and when unloaded. Electron-microscopic examination revealed processes of degeneration and lysis, suggesting apoptotic processes, in intracellular amastigotes and free amastigotes treated with the nanoparticles. It was demonstrated that unloaded nanoparticles, by mechanisms not completely elucidated, have trypanocide activity similar to that of a standard solution of nifurtimox. It is concluded that the nanoparticles loaded with nifurtimox constitutes a good carrier of the drug against T. cruzi. The loaded-nanoparticles significantly increase trypanocidal activity.